Phosphomimetics at Ser199/Ser202/Thr205 in Tau Impairs Axonal Transport in Rat Hippocampal Neurons

Christensen, Kyle R; Combs, Benjamin; Richards, Collin; Grabinski, Tessa; Alhadidy, Mohammed M; Kanaan, Nicholas M.

doi:10.1007/s12035-023-03281-3

Cited by 9 publications

(10 citation statements)

References 91 publications

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“…Modification mimicking approaches involve the substitution of specific amino acids with alternative amino acids that recapitulate the effects of a specific PTM. In the case of phosphorylation, serine and threonine residues are mutated to glutamate or aspartate to mimic the additional negative charge from phosphorylation [ 93 , 123 , 124 ] or to alanine to prevent phosphorylation [ 125 ]. In the case of acetylation, lysine residues are changed to glutamine to mimic the neutralization of charge induced by acetylation or to arginine to mimic a nonacetylated lysine [ 126 ].…”

Section: Methods To Produce Recombinant Tau Protein With Ptmsmentioning

confidence: 99%

“…Site-specific PTMs using mimetic approaches enriched our understanding of the roles PTMs play in regulating tau biology in health and disease. Pseudophosphorylation at either T175 or the S199/S202/T205 alters tau conformation leading to increased exposure of tau's PAD [ 51 , 93 , 98 ], which is a conformational change linked to tau-induced axonal transport impairment [ 64 , 93 ]. Moreover, pseudophosphorylation at S202 and T205 enhances the aggregation of tau in the presence of arachidonic acid [ 128 ], and modification of S199 or T205 alone is sufficient to impair axonal transport [ 129 ].…”

Section: Methods To Produce Recombinant Tau Protein With Ptmsmentioning

confidence: 99%

“…In conclusion, several lines of evidence demonstrate that PTMs of tau can change tau biology, leading to alterations in the levels of pathogenic conformations that are associated with neurotoxicity [ 93 , 97 , 98 ]. Subsequently, a better understanding of how specific PTMs affect tau aggregation, conformation and function is of high interest in the field.…”

Section: Introductionmentioning

confidence: 99%

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Biochemical approaches to assess the impact of post-translational modifications on pathogenic tau conformations using recombinant protein

Alhadidy,

Kanaan

2024

Biochemical Society Transactions

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Tau protein is associated with many neurodegenerative disorders known as tauopathies. Aggregates of tau are thought of as a main contributor to neurodegeneration in these diseases. Increasingly, evidence points to earlier, soluble conformations of abnormally modified monomers and multimeric tau as toxic forms of tau. The biological processes driving tau from physiological species to pathogenic conformations remain poorly understood, but certain avenues are currently under investigation including the functional consequences of various pathological tau changes (e.g. mutations, post-translational modifications (PTMs), and protein–protein interactions). PTMs can regulate several aspects of tau biology such as proteasomal and autophagic clearance, solubility, and aggregation. Moreover, PTMs can contribute to the transition of tau from normal to pathogenic conformations. However, our understating of how PTMs specifically regulate the transition of tau into pathogenic conformations is partly impeded by the relative lack of structured frameworks to assess and quantify these conformations. In this review, we describe a set of approaches that includes several in vitro assays to determine the contribution of PTMs to tau's transition into known pathogenic conformations. The approaches begin with different methods to create recombinant tau proteins carrying specific PTMs followed by validation of the PTMs status. Then, we describe a set of biochemical and biophysical assays that assess the contribution of a given PTM to different tau conformations, including aggregation, oligomerization, exposure of the phosphatase-activating domain, and seeding. Together, these approaches can facilitate the advancement of our understanding of the relationships between PTMs and tau conformations.

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Section: Methods To Produce Recombinant Tau Protein With Ptmsmentioning

confidence: 99%

Section: Methods To Produce Recombinant Tau Protein With Ptmsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Biochemical approaches to assess the impact of post-translational modifications on pathogenic tau conformations using recombinant protein

Alhadidy,

Kanaan

2024

Biochemical Society Transactions

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“…In vitro, co-incubation of tau aggregates with PP2A has been found to restore the binding of tau to microtubules to a level similar to the control group (approximately 80 %). Therefore, PP2A is the primary tau phosphatase, and its activators have promising potential for AD treatment [69,70].…”

Section: Phosphatase Activatormentioning

confidence: 99%

Targeting Abnormal Tau Phosphorylation for Alzheimer’s Therapeutics

Singh,

Ansari,

Mahmood

et al. 2024

Horm Metab Res

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Alzheimer’s disease (AD) is a widespread neurodegenerative disorder characterized by progressive memory and cognitive decline, posing a formidable public health challenge. This review explores the intricate interplay between two pivotal players in AD pathogenesis: β-amyloid (Aβ) and tau protein. While the amyloid cascade theory has long dominated AD research, recent developments have ignited debates about its centrality. Aβ plaques and tau NFTs are hallmark pathologies in AD. Aducanumab and lecanemab, monoclonal antibodies targeting Aβ, have been approved, albeit amidst controversy, raising questions about the therapeutic efficacy of Aβ-focused interventions. On the other hand, tau, specifically its hyperphosphorylation, disrupts microtubule stability and contributes to neuronal dysfunction. Various post-translational modifications of tau drive its aggregation into NFTs. Emerging treatments targeting tau, such as GSK-3β and CDK5 inhibitors, have shown promise in preclinical and clinical studies. Restoring the equilibrium between protein kinases and phosphatases, notably protein phosphatase-2A (PP2A), is a promising avenue for AD therapy, as tau is primarily regulated by its phosphorylation state. Activation of tau-specific phosphatases offers potential for mitigating tau pathology. The evolving landscape of AD drug development emphasizes tau-centric therapies and reevaluation of the amyloid cascade hypothesis. Additionally, exploring the role of neuroinflammation and its interaction with tau pathology present promising research directions.

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“…Another common method involves enzymatic functionalization of Tau to obtain phosphorylated, acetylated, ubiquitinated, and glycosylated forms, but this method lacks specificity and control over the location or extent of modifications. Furthermore, preparation of pure Tau is challenging because, as a disordered protein, Tau is vulnerable to protease degradation. , While more site-specific methods exist, they rely on the use of point mutations that mimic charge-neutralizing PTMs (S to D mutations for phosphoserine − or K to Q for acetyl lysine − ) or by using bioconjugate functionalization of dehydroalanine . The major disadvantage of this strategy is that these mimics may not possess the same structure, acidity, or charge as natural PTMs.…”

Section: Introductionmentioning

confidence: 99%

Chemical Synthesis of Microtubule-Associated Protein Tau

Powell,

Jing,

Walczak

2023

J. Am. Chem. Soc.

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Deposits of the microtubule-associated protein Tau (MAPT) serve as a hallmark of neurodegenerative diseases known as tauopathies. Numerous studies have demonstrated that in diseases such as Alzheimer's disease (AD), Tau undergoes extensive remodeling. The attachment of post-translational modifications distributed throughout the entire sequence of the protein correlates with clinical presentation. A systematic examination of these protein alterations can shed light on their roles in both healthy and diseased states. However, the ability to access these modifications in the entire protein chain is limited as Tau can only be produced recombinantly or through semisynthesis. In this article, we describe the first chemical synthesis of the longest 2N4R isoform of Tau, consisting of 441 amino acids. The 2N4R Tau was divided into 3 major segments and a total of 11 fragments, all of which were prepared via solid-phase peptide synthesis. The successful chemical strategy has relied on the strategic use of two cysteine sites (C291 and C322) for the native chemical ligations (NCLs). This was combined with modern preparative protein chemistries, such as mercaptothreonine ligation (T205), diselenide−selenoester ligation (D358), and mutations of mercaptoamino acids into native residues via homogeneous radical desulfurization (A40, A77, A119, A157, A246, and A390). The successful completion of the synthesis has established a robust and scalable route to the native protein in multimilligram quantities and high purity. In broader terms, the presented strategy can be applied to the preparation of other shorter isoforms of Tau as well as to introduce all post-translational modifications that are characteristic of tauopathies such as AD.

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Phosphomimetics at Ser199/Ser202/Thr205 in Tau Impairs Axonal Transport in Rat Hippocampal Neurons

Cited by 9 publications

References 91 publications

Biochemical approaches to assess the impact of post-translational modifications on pathogenic tau conformations using recombinant protein

Biochemical approaches to assess the impact of post-translational modifications on pathogenic tau conformations using recombinant protein

Targeting Abnormal Tau Phosphorylation for Alzheimer’s Therapeutics

Chemical Synthesis of Microtubule-Associated Protein Tau

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